Protein Synthesis ; Translation

TAU : An Innovative Centre of Excellence in Higher Medical Education & Research

Dr MOHAMED ABDELBAKY
Associate Professor
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 Learning Objectives
At the end of the lecture, students will be able to:
Study the genetic code and its characteristics.

Study in detail the process of translation (or

protein biosynthesis) and their inhibitors.

Study gene expression in prokaryotes and in

eukaryotes.

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Introduction
• Genetic information, stored in the chromosomes is
expressed through transcription to RNA (mRNA) and
subsequent translation into proteins.

• The process of protein synthesis is called translation

because the “language” of the nucleotide sequence on
the mRNA is translated into the language of an amino
acid sequence.

• The process of translation requires a genetic code,

through which the information contained in the nucleic
acid sequence is expressed to produce a specific
sequence of amino acids.
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Introduction
• Any change in the nucleic acid sequence may
result in an improper amino acid of the protein
chain may causing disease or even death of the
organism.

• Many proteins are covalently modified following

their synthesis to activate them or change their
activities.

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 The Genetic Code
o The genetic code is a dictionary that identifies
the correspondence between a sequence of
nucleotide bases on mRNA and a sequence of
amino acids.

o Each word in the code is composed of three

nucleotide bases.

o These genetic words are called codons.

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A.Codons
 Codons are presented in the messenger RNA (mRNA)
language of adenine (A), guanine (G), cytosine (C), and
uracil (U).

 Their nucleotide sequences are always written from the

5′ end to the 3′ end.

 The four nucleotide bases are used to form the three-

base codons.

 There are 64 different combinations of bases when

three nucleotide taken at a time.
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1) Translation of the Codon:
 The “dictionary” can be used to translate any
codon sequence to determine which amino
acids are coded for by an mRNA sequence.

 For example, the codon 5′-AUG-3′ codes for

methionine.

 61 of the 64 codons code for the 20 common

amino acids.

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2) Termination (“stop” or “nonsense”)
codons:
 Three of the codons, UAG, UGA, and UAA, do
not code for amino acids but they are
termination codons.

 When one of these codons appears in an mRNA

sequence, it signals that the synthesis of the
protein by that mRNA is complete.

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B. Characteristics of the Genetic Code
 The genetic code is consistent throughout all
living organisms.

 Characteristics of the genetic code include the

following:

1. Specificity:
The genetic code is specific (unambiguous), that is,
a particular codon always codes for the same
amino acid.
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2. Universality:
 The genetic code is virtually universal, that is, the
specificity of the genetic code has been conserved
from very early stages of evolution (with slight
differences) in the manner in which that code is
translated.

 Note: An exception occurs in mitochondria, in which a

few codons are different e.g., UGA codes for trp while
in nuclear DNA and its mRNA is a stop codon.

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3. Degeneracy:
 The genetic code is degenerate (redundant).

 Although each codon corresponds to a single

amino acid, a given amino acid may have
more than one triplet coding for it.

 For example, arginine is coded by six different

codons.

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4. Nonoverlapping and commaless:
• The genetic code is read without overlapping
and commas, that is, the code is read from a
fixed starting point as a continuous sequence
of three bases, taken at a time.

• For example, AUGCCUGGAUAA is read as

AUG/CCU/GGA/UAA without any “overlap or
punctuation” between the codons.

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C. Alteration in the Nucleotide
Sequence
Point mutation is a single nucleotide base change on the
mRNA chain can lead to any one of three results:

1)Silent Mutation
oThe codon containing the new changed base may
specifies for the same amino acid.

o For example, the serine codon UCA when changed to

UCU, it still codes for serine and this is termed a silent
mutation.

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2) Missense Mutation
o The changed codon containing the new base may code
for a different amino acid.

o For example, if the serine codon UCA is changed to

become CCA, it will code for a different amino acid, in
this case, proline.

o The substitution of an incorrect amino acid is called a

missense mutation.

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3) Nonsense Mutation
 The codon containing the changed base may become a
termination or stop codon.

 For example, if the serine codon UCA is changed in a

different second base “A” and to become UAA, the
new codon causes termination of translation at that
point and this leads to the production of a shortened
(truncated) protein.

 The creation of a termination codon or stop codon at

an inappropriate place is called a nonsense mutation.
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 Components Required For Translation
A large number of components are required for the
synthesis of a protein, these include:
1.All the amino acids
2.The mRNA to be translated
3.Transfer RNA (tRNA)
4.Functional ribosomes
5.Energy sources
6.Enzymes, as well as protein factors needed for
initiation, elongation, and termination of the
polypeptide chain.
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A. Amino Acids
 All the amino acids that should be appear in the
complete protein must be present at the time of
protein synthesis.
 If one amino acid is missing [e.g., if the diet does
not contain an essential amino acid],
translation stops at the codon specifying that
amino acid.
 It is important to having all the essential amino
acids in sufficient quantities in the diet to ensure
continued protein synthesis.
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B. Transfer RNA
• tRNAs are able to recognize the codon on mRNA for a
amino acid and carry the specific amino acid and so
tRNA functions as adaptor molecules.

• At least one specific type of tRNA is required per amino

acid.

• In humans, there are at least 50 species of tRNA,

whereas bacteria contain 30 to 40 species.

• Because there are only 20 different amino acids, some

amino acids have more than one specific tRNA
molecule (amino acids that are coded for by several
codons). 23
i. tRNA Amino Acid Attachment Site:

• Each tRNA molecule has an attachment site for a

specific amino acid at its 3′ end; the carboxyl group of
the amino acid is linked with an ester linkage with the
3′-hydroxyl group of the ribose moiety of the adenosine
nucleotide in the –CCA sequence at the 3′ end of the
tRNA.

• When a tRNA has a covalently attached amino acid, it is

said to be charged; when tRNA is not bound to an
amino acid, it is described as being uncharged.

• The amino acid that is attached to the tRNA molecule,

it is called activated.
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ii. tRNA Anticodon:
o Each tRNA molecule also contains a three-base
nucleotide sequence (the anticodon) that
recognizes a specific codon on the mRNA.

o This codon of the mRNA specifies the insertion

of the amino acid into the growing peptide chain
carried by that tRNA.

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C. Aminoacyl-tRNA Synthetases
o A family of enzymes is required for the attachment of
amino acids to their corresponding tRNA.

o Each enzyme member of this family recognizes a

specific amino acid and the tRNA that corresponds to
that amino acid (isoaccepting tRNA).

o These enzymes can read both the three-letter code of

nucleic acids of tRNA and the 20-letter code of amino
acids.

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o Each aminoacyl-tRNA synthetase catalyzes a two-step
reaction that results in the covalent attachment of the
carboxyl group of an amino acid to the 3′ end of its
corresponding tRNA.

o The reaction needs adenosine triphosphate (ATP),

which is cleaved to adenosine monophosphate (AMP)
and inorganic pyrophosphate (PPi).

o The synthetase has extreme specificity in recognizing

both the amino acid and its specific tRNA contributes to
the high fidelity of translation of the genetic message.
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o In addition, the synthetases have a proofreading or
editing activity it can remove mischarged amino acids
from the enzyme or from the tRNA molecule.

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D. Functionally Competent Ribosomes
 Ribosomes are large complexes of protein and
ribosomal RNA (rRNA).
 They consist of two subunits; one large and one small
and relative sizes are generally given in terms of their
sedimentation coefficients, or S (Svedberg) values.
 The S values are determined both by shape as well as
by molecular mass and so their numeric values are not
strictly additive.
 The eukaryotic 60S and 40S subunits form an 80S
ribosome.
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 Prokaryotic and eukaryotic ribosomes are similar in
structure and have the same function as the factories
of proteins synthesis.

 The smaller subunit binds mRNA and is responsible for

the accuracy of translation by allow correct base-
pairing between the codon in the mRNA and the
anticodon of the tRNA.

 The larger ribosomal subunit catalyzes the formation of

the peptide bonds that link amino acid residues in a
protein.
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A, P, and E Sites on the Ribosome:
 The ribosome has three binding sites for tRNA
molecules; the A, P, and E sites.

 They cover three neighboring codons.

 During translation, the A site binds an incoming

aminoacyl-tRNA as directed by the codon currently
occupying this site.

 The P-site codon is occupied by peptidyl-tRNA.

 The E site is occupied by the empty tRNA as it is about

to exit the ribosome. 34
 A site = incoming Aminoacyl-tRNA.
 P site = accommodates growing Peptide.
 E site = holds Empty tRNA as it Exits.

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E. Protein Factors
 Initiation, elongation, and termination (or
release) factors are required for peptide
synthesis.

 Some of these protein factors have a catalytic

function, whereas others appear to stabilize the
synthetic machinery of polypeptide.

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F. ATP and GTP as Sources of Energy
 Four high-energy bonds is required for the
addition of one amino acid to the growing
polypeptide chain: two from ATP in the
aminoacyl-tRNA synthetase reaction and two
from GTP; one for binding the aminoacyl-tRNA
to the A site and one for the translocation step.

 In eukaryotes, additional ATP and GTP

molecules are required for initiation and an
additional GTP molecule is required for
termination.
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 Codon Recognition by tRNA
 Correct pairing of the codon in the mRNA with
the anticodon of the tRNA is essential for
accurate translation.

 Some tRNAs recognize more than one codon for

a given amino acid.

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I. Antiparallel Binding Between Codon and
Anticodon:
 Binding of the tRNA anticodon to the mRNA codon
follows the rules of complementary and antiparallel
binding.

 The mRNA codon is “read” 5′ → 3′ is pairing to an

anticodon in the “flipped” (3′ → 5′) orientation.

 When writing the sequences of both codons and

anticodons, the nucleotide sequence must always be
listed in the 5′ → 3′ order.

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II. Wobble Hypothesis
• The mechanism by which tRNAs can recognize more
than one codon for a specific amino acid is described by
the “wobble hypothesis” in which the base at the 5′
end of the anticodon (the first base of the anticodon)
is not importantly identified as the other two bases.

• This allows nontraditional base-pairing between the 3′

base of the codon (last base of the codon).

• This “wobble hypothesis” allows a single tRNA to

recognize more than one codon

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• Wobble: codons that differ in 3rd (“wobble”) position may code
for the same tRNA/amino acid.

• Specific base pairing is usually required only in the first 2

nucleotide positions of mRNA codon.

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 Textbook/ Course Reference
1) Genetics in Medicine, Thompson & Thompson, 8th
Edition, 2016, ISBN-13: 978-1437706963.
2) Medical Genetics, Ian D. Young, ISBN-13:
9780199594610 2010.
3) Board Review Series (BRS)-GENETICS: Ronald W
Derduk- Published by Lippincott-Williams and Wilkins,
ISBN-13: 978-0781799942, 2013.
4) Biochemistry and Genetics PreTest.
5) Lippincott's Illustrated Reviews: Cell and Molecular
Biology.

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 Questions
1. Polysomes do not contain:
A. Protein
B. DNA
C. m-RNA
D. t-RNA

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1. Polysomes do not contain:
A. Protein
B. DNA
C. m-RNA
D. t-RNA

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2. The formation of a peptide bond during the
elongation step of protein synthesis results in
the splitting of how many high energy bonds?
A. 1
B. 3
C. 4
D. 2

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2. The formation of a peptide bond during the
elongation step of protein synthesis results in
the splitting of how many high energy bonds?
A. 1
B. 3
C. 4
D. 2

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3. Translocase is an enzyme required in the
process of:
A. DNA replication
B. RNA synthesis
C. Initiation of protein synthesis
D. Elongation of peptides

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3. Translocase is an enzyme required in the
process of:
A. DNA replication
B. RNA synthesis
C. Initiation of protein synthesis
D. Elongation of peptides

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4. Nonsense codons bring about:
A. Amino acid activation
B. Initiation of protein synthesis
C. Termination of protein synthesis
D. Elongation of polypeptide chains

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4. Nonsense codons bring about:
A. Amino acid activation
B. Initiation of protein synthesis
C. Termination of protein synthesis
D. Elongation of polypeptide chains

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5. The anticodon region is an important part of
the structure of:
A. r-RNA
B. t-RNA
C. m-RNA
D. hn-RNA

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5. The anticodon region is an important part of
the structure of:
A. r-RNA
B. t-RNA
C. m-RNA
D. hn-RNA

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6. Degeneracy of the genetic code denotes the
existence of:
A. Base triplets that do not code for any amino
acids
B. Codons consisting of only two bases
C. Codons that include one or more unusual bases
D. Multiple codons for a single amino acid

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6. Degeneracy of the genetic code denotes the
existence of:
A. Base triplets that do not code for any amino
acids
B. Codons consisting of only two bases
C. Codons that include one or more unusual bases
D. Multiple codons for a single amino acid

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